LysoBrite™ Blue
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Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Molecular weight | 331.42 |
Solvent | DMSO |
Excitation (nm) | 434 |
Emission (nm) | 480 |
Certificate of Origin | Download PDF |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
Storage | Freeze (< -15 °C); Minimize light exposure |
UNSPSC | 12352200 |
LysoBrite™ Red DND-99 |
LysoBrite™ Green DND-26 |
Overview | SDSProtocol |
Molecular weight 331.42 | Excitation (nm) 434 | Emission (nm) 480 |
Platform
Flow cytometer
Excitation | 405 nm laser |
Emission | 450/40 nm filter |
Instrument specification(s) | Pacific Blue channel |
Fluorescence microscope
Excitation | DAPI filter set |
Emission | DAPI filter set |
Recommended plate | Black wall/clear bottom |
Example protocol
AT A GLANCE
Prepare cells.
Add dye working solution.
Incubate at 37 °C for 30 minutes.
Wash the cells.
Analyze under a fluorescence microscope.
The LysoBrite™ Blue stock solution provided is 500X in DMSO. It should be stable for at least 6 months if stored at -20°C. Protect from light, and avoid freeze/thaw cycles.
PREPARATION OF WORKING SOLUTION
Warm LysoBrite™ Blue dye to room temperature.
Prepare dye working solution by diluting 20 µL of 500X LysoBrite™ Blue with 10 mL of Hanks and 20 mM HEPES buffer (HBSS) or buffer of your choice.
Note: 20 µL of LysoBrite™ Blue dye is enough for one 96-well plate. Aliquot and store unused LysoBrite™ dye stock solutions at < -15 °C. Protect it from light and avoid repeated freeze-thaw cycles.
Note: The optimal concentration of the fluorescent lysosome indicator varies depending on the specific application. The staining conditions may be modified according to the particular cell type and the permeability of the cells or tissues to the probe.
SAMPLE EXPERIMENTAL PROTOCOL
This protocol only provides a guideline and should be modified according to your specific needs.
Grow cells in a 96-well black wall/clear bottom plate (100 µL/well/96-well plate) or on coverslips inside a petri dish filled with the appropriate culture medium.
When cells reach the desired confluence, add an equal volume of the dye-working solution (from Preparation of Working Solution Step 2).
Incubate the cells in a 37 °C, 5% CO2 incubator for 30 minutes.
Wash the cells twice with pre-warmed (37 °C) Hanks and 20 mM HEPES buffer (HBSS) or buffer of your choice. Then fill the cell wells with HBSS or growth medium.
Observe the cells using a fluorescence microscope fitted with the desired filter set.
Note: It is recommended to increase either the labeling concentration or the incubation time to allow the dye to accumulate if the cells do not appear to be sufficiently stained.
Add an equal volume of the dye-working solution (from Preparation of Working Solution Step 2).
Incubate the cells in a 37 °C, 5% CO2 incubator for 30 minutes.
Wash the cells twice with pre-warmed (37 °C) Hanks and 20 mM HEPES buffer (HBSS) or buffer of your choice. Then fill the cell wells with HBSS or growth medium.
Observe the cells using a fluorescence microscope fitted with the desired filter set.
Note: It is recommended to increase either the labeling concentration or the incubation time to allow the dye to accumulate if the cells do not appear to be sufficiently stained.
Note: Suspension cells may be attached to coverslips treated with BD Cell-Tak® (BD Biosciences) and stained as adherent cells (see Protocol for Preparing and Staining Adherent Cells).
Calculators
Common stock solution preparation
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 301.732 µL | 1.509 mL | 3.017 mL | 15.087 mL | 30.173 mL |
5 mM | 60.346 µL | 301.732 µL | 603.464 µL | 3.017 mL | 6.035 mL |
10 mM | 30.173 µL | 150.866 µL | 301.732 µL | 1.509 mL | 3.017 mL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
/ | = | x | = |
Product Family
Name | Excitation (nm) | Emission (nm) |
LysoBrite™ Green | 501 | 510 |
LysoBrite™ Orange | 543 | 565 |
LysoBrite™ Red | 576 | 596 |
LysoBrite™ Deep Red | 597 | 619 |
LysoBrite™ NIR | 636 | 651 |
Images
Citations
Authors: Wei, Dan and Wang, Luolin and Lei, Shunmei and Zhang, Han and Dong, Caihua and Ke, Yao and Su, Yuting and Chen, Xiaoying and Xia, Lianping and Kong, Xiaoyang and others,
Journal: Pharmaceutical Biology (2023): 839--857
Authors: Fan, Shuangqi and Wu, Keke and Zhao, Mingqiu and Yuan, Jin and Ma, Shengming and Zhu, Erpeng and Chen, Yuming and Ding, Hongxing and Yi, Lin and Chen, Jinding
Journal: Autophagy (2020): 1--20
Authors: Zhang, Xiao and Fan, Jiabing and Lee, Chung-Sung and Kim, Soyon and Chen, Chen and Aghaloo, Tara and Lee, Min
Journal: Advanced Functional Materials (2020): 1909218
Authors: Hou, Ji-Ting and Kim, Hyeong Seok and Duan, Chong and Ji, Myung Sun and Wang, Shan and Zeng, Lintao and Ren, Wen Xiu and Kim, Jong Seung
Journal: Chemical communications (2019): 2533--2536
Authors: Lin, Yi and Yang, Yidi and Yan, Jianqin and Chen, Jun and Cao, Jun and Pu, Yuji and Li, Li and He, Bin
Journal: Journal of Materials Chemistry B (2018): 2089--2103
Authors: Yuan, Xuexia and Xiao, Fan and Zhao, Haoran and Huang, Yishun and Shao, Chen and Tian, Leilei and Weizmann, Yossi
Journal: ACS Applied Bio Materials (2018)
Authors: Cemma, Marija and Grinstein, Sergio and Brumell, John H
Journal: Autophagy (2016): 1440--1446
Authors: Myung, Ja Hye and Gajjar, Khyati A and Chen, Jihua and Molokie, Robert E and Hong, Seungpyo
Journal: Analytical chemistry (2014): 6088--6094
Authors: Lei, Bo and Chen, Xiaofeng and Han, Xue and Zhou, Jiaan
Journal: Journal of Materials Chemistry (2012): 16906--16913
References
Authors: Gough AH, Johnston PA.
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Journal: Methods Mol Biol (2007): 189
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Journal: Methods Mol Biol (2007): 3
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Journal: Methods Enzymol (2006): 21
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Journal: Assay Drug Dev Technol (2006): 209
Authors: Hudson CC, Oakley RH, Sjaastad MD, Loomis CR.
Journal: Methods Enzymol (2006): 63
Authors: Werner T, Liebisch G, Gr and l M, Schmitz G.
Journal: Cytometry A (2006): 200
Authors: Wolff M, Haasen D, Merk S, Kroner M, Maier U, Bordel S, Wiedenmann J, Nienhaus GU, Valler M, Heilker R.
Journal: Comb Chem High Throughput Screen (2006): 339
Authors: O'Brien P J, Irwin W, Diaz D, Howard-Cofield E, Krejsa CM, Slaughter MR, Gao B, Kaludercic N, Angeline A, Bernardi P, Brain P, Hougham C.
Journal: Arch Toxicol (2006): 580
Application notes
Fluorescent Oligonucleotide Labeling Reagents
Monitoring of Mitochondrial Membrane Potential Changes in Live Cells Using JC-10
Selective Analysis of RNA in Live and Fixed Cells with StrandBrite RNA Green
Cell Loading Protocol For Fluorescent pH Indicator, BCECF-AM
FAQ
I ordered your phalloidin-amine (Cat #5302) so I can conjugate it to my oligo. Do you have a recommended protocol I can use?
How can I lyse my cells without lysing the nuclear membrane?
Do you have any dual-fluorescence nucleic acid stains that interact with both DNA and RNA?
Do you have any fixable mitochondria staining assay kits?